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目的:了解肺组织内被严重急性呼吸综合征冠状病毒(severeacutere spiratorysyndromeas sociatedcoron avirus,SARS CoV)感染的靶细胞类型,并对SARS诱发的肺损伤发病机制进行探讨。方法:运用SARSCoV基因组序列合成的地高辛标记cDNA探针,对北京市7例及安徽省1例确诊的SARS死亡病例的肺组织进行原位杂交检测,在原位杂交基础上,进一步运用免疫组织化学方法显示SARSCoV感染的靶细胞类型,如Cytokeratin(CK)标记上皮细胞,CD34标记血管内皮细胞,CD68标记巨噬细胞,Vimentin标记纤维母细胞,CD3标记全T细胞。结果:原位杂交检测显示,8例患者肺组织中都表达SARSCoVRNA,阳性信号位于靶细胞胞浆内,呈紫蓝色(NBTBCIP)。原位杂交和免疫组化结果显示:支气管上皮细胞、Ⅱ型肺泡上皮细胞、血管内皮细胞、巨噬细胞、纤维母细胞及T淋巴细胞在所有SARS病例中都受到了病毒感染。原位杂交阳性(紫蓝色,NBTBCIP)和免疫组化阳性(红棕色,AEC)信号同时表达于靶细胞胞浆中而呈紫红色。结论:通过对SARS患者肺组织原位杂交和免疫组织化学双重标记研究表明,肺组织内支气管上皮细胞、Ⅱ型肺泡上皮细胞、血管内皮细胞、巨噬细胞及纤维母细胞等多种细胞成分广泛受到了SARSCoV攻击,肺组织内多种细胞成分弥漫性受损以及所释放的炎性介质在肺损伤的发病过程中起着重要作用。
Objective: To know the types of target cells in lung tissue infected by severe acute respiratory syndrome virus (SARS CoV) and to explore the pathogenesis of SARS-induced lung injury. Methods: Using digoxigenin-labeled cDNA probe synthesized by SARSVV genome sequence, in situ hybridization was detected in 7 cases of SARS in Beijing and 1 case of SARS death in Anhui Province. Based on the in situ hybridization, further immunization Histochemical methods revealed the target cell types of SARSCoV infection, such as Cytokeratin (CK) labeled epithelial cells, CD34 labeled vascular endothelial cells, CD68-labeled macrophages, Vimentin-labeled fibroblasts and CD3-labeled whole T cells. Results: In situ hybridization showed that SARSCoVRNA was expressed in lung tissues of 8 patients. The positive signal was located in the cytoplasm of target cells, showing a purple-blue color (NBTBCIP). In situ hybridization and immunohistochemistry results showed that bronchial epithelial cells, type II alveolar epithelial cells, vascular endothelial cells, macrophages, fibroblasts and T lymphocytes were all infected with virus in all SARS cases. In situ hybridization positive (purple blue, NBTBCIP) and immunohistochemical (red brown, AEC) signals were expressed in the cytoplasm of the target cells with a purplish red color. Conclusion: In situ hybridization and immunohistochemical double labeling of lung tissue in SARS patients showed that a wide range of cellular components such as bronchial epithelial cells, type II alveolar epithelial cells, vascular endothelial cells, macrophages and fibroblasts in lung tissues SARSCoV attack, diffuse damage to a variety of cellular components in the lung tissue and the release of inflammatory mediators play an important role in the pathogenesis of lung injury.